Chapman M D, Sutherland W M, Platts-Mills T A
J Immunol. 1984 Nov;133(5):2488-95.
Monoclonal antibodies were raised against Antigen P1, the major allergen of the house dust mite (Dermatophagoides pteronyssinus). The majority were Antigen P1 specific, isotype IgG1, and did not react with a comparable D. farinae allergen. These antibodies bound 38 to 50% of 125I Antigen P1 in antigen-binding assays (titer greater than or equal to 1/1,000,000), and the quantities of IgG antibody in ascites were 2 to 4 logs greater than those in polyclonal mouse antiserum or in serum from a mite-allergic patient. Two IgM antibodies showed weak binding to Antigen P1 but reacted strongly with D. pteronyssinus in enzyme immunoassay (titer greater than or equal to 1/100,000). Assessments of the specificity of the IgG antibodies by using two inhibition radioimmunoassays suggested that they were directed against two different epitopes. Antibodies 10B9 F6 and 5H8 C12 were purified by preparative isoelectric focusing (isoelectric points of pI 6.25 and 7.4, respectively) and radiolabeled with 125I. Cross-inhibition experiments, using ascites dilutions to inhibit binding of each radiolabeled antibody to Antigen P1, confirmed that these antibodies recognized two distinct epitopes. Analysis of antibodies from 39 clones/hybrids showed that the majority were directed against the same epitopes as either 10B9 F6 or 5H8 C12 (3 out of 39 [8%] and 29 out of 39 [74%], respectively). None of the monoclonal antibodies significantly inhibited (greater than 10%) human IgE binding to Antigen P1 in the radioallergosorbent test. However, 12 of 14 sera from mite allergic patients inhibited binding by the monoclonal antibodies. One serum from a mite-allergic patient inhibited binding of both 10B9 F6 and 5H8 C12 by greater than 85% and showed parallel inhibition curves. The results suggest that these monoclonal antibodies could be used to assay Antigen P1 in both D. pteronyssinus and house dust extracts. It should also be possible to use monoclonal antibodies in inhibition assays to define the antigenic/allergenic determinants recognized by human IgG and IgE antibodies on this mite allergen.
制备了针对屋尘螨（粉尘螨）主要变应原抗原P1的单克隆抗体。大多数抗体具有抗原P1特异性，为IgG1同种型，且不与粗脚粉螨的类似变应原发生反应。在抗原结合试验中，这些抗体可结合38%至50%的125I标记抗原P1（效价大于或等于1/1,000,000），腹水IgG抗体量比多克隆小鼠抗血清或螨过敏患者血清中的IgG抗体量高2至4个对数级。两种IgM抗体与抗原P1的结合较弱，但在酶免疫测定中与粉尘螨反应强烈（效价大于或等于1/100,000）。使用两种抑制放射免疫测定法评估IgG抗体的特异性，结果表明它们针对两个不同的表位。抗体10B9 F6和5H8 C12通过制备性等电聚焦进行纯化（等电点分别为pI 6.25和7.4），并用125I进行放射性标记。使用腹水稀释液抑制每种放射性标记抗体与抗原P1结合的交叉抑制实验证实，这些抗体识别两个不同的表位。对39个克隆/杂交体的抗体分析表明，大多数抗体针对与10B9 F6或5H8 C12相同的表位（分别为39个中的3个[8%]和39个中的29个[74%]）。在放射变应原吸附试验中，没有一种单克隆抗体能显著抑制（大于10%）人IgE与抗原P1的结合。然而，14份螨过敏患者血清中的12份可抑制单克隆抗体的结合。一份螨过敏患者的血清可抑制10B9 F6和5H8 C12的结合达85%以上，并呈现平行抑制曲线。结果表明，这些单克隆抗体可用于检测粉尘螨和屋尘提取物中的抗原P1。也应该能够在抑制试验中使用单克隆抗体来确定人IgG和IgE抗体在此螨变应原上识别的抗原/变应原决定簇。
